RAR PROTAC compound, pharmaceutical composition, preparation method and application

Abstract

A retinoic acid receptor RAR PROTAC molecule with a novel structure is designed and synthesized, and RAR is degraded in a targeted manner by utilizing a PROTAC technology, so that the compound has a very good inhibition effect on tumor stem cells, has good properties such as pharmacokinetics, bioavailability and in-vivo drug effect, and can be developed into a novel anti-tumor drug.

Description

Technical Field

[0001] The present invention belongs to the technical field of drug synthesis and application, and specifically relates to a RAR PROTAC compound, a pharmaceutical composition, a preparation method and an application. Background Art

[0002] Cancer stem cells (CSCs) are a special subpopulation of tumor cells that play a key role in the development, invasion and distant metastasis of tumors. CSCs have the ability to self-renew indefinitely or for a long time, which means that they can continuously divide and produce new tumor cells. Although CSCs may remain undifferentiated in the tumor microenvironment, they have the ability to differentiate into various types of tumor cells. CSCs are generally highly resistant to conventional chemotherapy and radiotherapy, and can also promote immune escape by affecting immune cells in the tumor microenvironment, which allows them to survive after treatment and may cause tumor recurrence and metastasis.

[0003] Drug development targeting CSCs is an important research direction. If CSCs can be effectively targeted and eliminated, the success rate of cancer treatment can be significantly improved and recurrence can be reduced. WYC-209, as a new type of synthetic retinoic acid drug, has shown significant inhibitory effects on CSCs in various types of tumors by targeting retinoic acid receptors (RAR), which provides new possibilities for the development of therapeutic strategies targeting CSCs.

[0004] Proteolysis-Targeting Chimeras (PROTACs) technology is a new protein degradation strategy that uses the intracellular ubiquitin-proteasome system (UPS) to specifically degrade target proteins. The core idea of ​​this technology is to design and develop small molecule compounds that can bind to two proteins at the same time: one is the target protein (POI) and the other is the E3 ubiquitin ligase; through this bispecific binding, PROTACs promote the ubiquitination of the target protein, thereby marking the protein for recognition and degradation by the proteasome, which makes PROTACs show great potential in the treatment of tumor-induced diseases. In view of this, using PROTAC technology to develop a series of PROTAC molecules that target the degradation of RAR will be a potential targeted therapeutic strategy for inhibiting malignant tumor stem cells. Summary of the invention

[0005] The purpose of the present invention is to provide a RAR PROTAC compound that can at least solve some of the defects existing in the prior art.

[0006] To achieve the above object, the present invention adopts the following technical solution:

[0007] A RAR PROTAC compound has a structure shown in the following general formula:

[0008]

[0009] Among them, E3 ligand is selected from IAP ligand

[0010]

[0011] One of them.

[0012] Furthermore, the Linker is a rigid linker, a flexible linker, or a flexible and rigid combination linker.

[0013] Furthermore, the rigid linking group is selected from one of the following structural formulas:

[0014]

[0015] The flexible linking group is selected from one of the following structural formulas:

[0016]

[0017]

[0018] The flexible and rigid combined linking group is selected from one of the following structural formulas:

[0019]

[0020] Among them, n=0-10.

[0021] Further, the RAR PROTAC compound is selected from any one of the following compounds:

[0022]

[0023]

[0024]

[0025]

[0026] In addition, the present invention also provides a method for preparing the above-mentioned RAR PROTAC compound, comprising the following steps:

[0027] S1. Synthesis of ethyl 2-((4,4-dimethyl-1-oxobenzothiopyran-6-)ethynyl)-4-hydroxypyrimidine-5-carboxylate, or synthesis of ethyl 2-((4,4-dimethyl-1-oxobenzothiopyran-6-)ethynyl)-4-bromopyrimidine-5-carboxylate;

[0028] S2. Using 2-((4,4-dimethyl-1-oxobenzothiopyran-6-)ethynyl)-4-hydroxypyrimidine-5-carboxylic acid ethyl ester or 2-((4,4-dimethyl-1-oxobenzothiopyran-6-)ethynyl)-4-bromopyrimidine-5-carboxylic acid ethyl ester synthesized in step S1 to prepare a retinoic acid compound;

[0029] S3, reacting the E3 ligand with the linker to obtain an intermediate compound;

[0030] S4. Prepare RAR PROTAC compounds using retinoic acid compounds and intermediate compounds.

[0031] Furthermore, in step S4, the process of preparing the RAR PROTAC compound from the retinoic acid compound and the intermediate compound is as follows: the retinoic acid compound and the intermediate compound are dissolved in N,N dimethylformamide, and then sodium triacetylborohydride or HATU is added, and the reaction is stirred at room temperature; after the reaction is completed, a saturated ammonium chloride solution is added to quench the reaction, and then ethyl acetate and water are added for extraction, the ethyl acetate phase is washed with saturated brine, and the reduced pressure distillation is followed by plate separation to obtain the RAR PROTAC compound.

[0032] The present invention also provides the use of the above-mentioned RAR PROTAC compound and a pharmaceutically acceptable salt thereof in the preparation of an anti-tumor drug.

[0033] Further, the pharmaceutically acceptable salt of the RAR PROTAC compound is a salt of the RAR PROTAC compound and any one of the following acids: hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, carbonic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, malic acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, succinic acid, fumaric acid, salicylic acid, phenylacetic acid, mandelic acid or ferulic acid.

[0034] Furthermore, the tumor is melanoma, breast cancer or prostate cancer.

[0035] The present invention also provides a pharmaceutical composition, comprising the above-mentioned RAR PROTAC compound, and one or more combinations of pharmaceutically acceptable carriers, excipients, diluents, adjuvants, and vehicles for the RARPROTAC compound.

[0036] Compared with the prior art, the present invention has the following beneficial effects:

[0037] The present invention designs and synthesizes a new structure of retinoic acid receptor RAR PROTAC molecules, uses PROTAC technology to target and degrade RAR, has a good inhibitory effect on tumor stem cells, and has good properties such as pharmacokinetics, bioavailability and in vivo efficacy, and can be developed into a new anti-tumor drug. DETAILED DESCRIPTION

[0038] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0039] The compounds of the present invention may exist in specific geometric or stereoisomeric forms. The present application contemplates all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (Z)-isomers and racemic mixtures thereof and other mixtures, such as enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of the present application. Additional asymmetric carbon atoms may be present in substituents such as alkyl. All of these isomers and their mixtures are included within the scope of the present application.

[0040] The compounds and intermediates of the present invention may also exist in different tautomeric forms, and all such forms are included within the scope of the present application. The term "tautomer" or "tautomeric form" refers to structural isomers of different energies that can interconvert via a low energy barrier. The compounds of the present application may contain unnatural proportions of atomic isotopes on one or more of the atoms that make up the compound. For example, compounds may be labeled with radioactive isotopes, such as tritium ( 2 H), iodine-125( 125 I) or C-14( 14 C). For another example, deuterium can be substituted for hydrogen to form a deuterated drug. All isotopic composition changes of the compounds of the present application, whether radioactive or not, are included in the scope of the present application.

[0041] The term "targeting chimera" refers to a bifunctional molecule comprising two small molecule ligands, one with high affinity for a target protein of interest, and a second for recruiting an E3 ligase that ubiquitinates the protein and targets it for proteolysis by the proteasome.

[0042] In the present invention, the structure of the target compound was determined by high resolution mass spectrometry (HR-ESI-MS), nuclear magnetic resonance spectroscopy ( 1 H-NMR,13 C-NMR) confirmation. NMR was determined using a Bruker Avance III 400 nuclear magnetic spectrometer, with deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), deuterated methanol (CD3OD), and tetramethylsilane (TMS) as the internal standard; MS was determined using an Agilent 6120B (ESI); HPLC was determined using an Agilent 1260DAD high performance liquid chromatograph (Zorbax SB-C18100×4.6mm, 3.5μM); thin layer chromatography silica gel plates used Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plates, the silica gel plates used for thin layer chromatography (TLC) used specifications of 0.15mm-0.20mm, and the specifications used for thin layer chromatography separation and purification products used specifications of 0.4mm-0.5mm; Yantai Huanghai silica gel 200-400 mesh silica gel was generally used as the carrier for column chromatography

[0043] The present invention provides a RAR PROTAC compound having a structure shown in the following general formula:

[0044]

[0045] Wherein, the E3 ligand is selected from IAP ligands

[0046]

[0047] One of them.

[0048] The linker can be selected from a rigid linker, a flexible linker, or a flexible and rigid combination linker. Specifically, the rigid linker is selected from one of the following structural formulas:

[0049]

[0050]

[0051] The flexible linking group is selected from one of the following structural formulas:

[0052]

[0053] in,

[0054] n=0-10.

[0055] The flexible and rigid combined linking group is selected from one of the following structural formulas:

[0056]

[0057] Among them, n=0-10.

[0058] Example 1: Synthesis of ethyl 2-((4,4-dimethylbenzothiopyran-6-)ethynyl)-4-hydroxypyrimidine-5-carboxylate (Compound 3)

[0059]

[0060] 6-Ethynyl-4,4-dimethylthiobenzopyran (i.e., compound 1) and ethyl 2-chloro-4-hydroxy-5-pyrimidinecarboxylate (i.e., compound 2) were added to a flask, and Pd(PPh3)2Cl2 (dichlorobis(triphenylphosphine)palladium) and CuI (copper iodide) were added. The flask was protected by argon and the gas was replaced three times to exclude oxygen. 5 ml of dry THF (anhydrous tetrahydrofuran) and dried Et3N (triethylamine) were added with a syringe, and the reaction was refluxed. After the reaction was completed as monitored by TLC thin-layer chromatography, THF was evaporated, ethyl acetate was added for extraction, and the mixture was washed with 5.0 eq 2N dilute hydrochloric acid, 2×30 mL saturated sodium bicarbonate solution and 30 mL saturated sodium chloride solution, respectively. The organic layer was dried over anhydrous sodium sulfate, filtered and dried to obtain a crude product, which was purified by column chromatography to obtain compound 3.

[0061] The test results of the product prepared above are as follows: 1 H NMR (400MHz, CDCl3) δ7.81(d,J=8.1Hz,1H),7.47(dd,J=6.2,2.2Hz,1H),7.36(s,0H),7.16(dd,J=8.1,2.1Hz,1H),7.09(d ,J=6.2Hz,1H),7.03(d,J=2.1Hz,1H),4.30(q,J=6.4Hz,2H),3.02-2.96(m,2H),2.07-2.01(m,2H),1.37(t,J=6.4Hz,3H). 13 C NMR (125 MHz, Common NMR Solvents) δ 170.08, 164.28, 143.59, 135.91, 131.59, 130.85, 128.73, 127.85, 126.37, 123.09, 120.70, 116.74, 115.41, 89.18, 61.42, 37.38, 37.09, 30.22, 26.90, 14.32; this indicated that compound 3 was synthesized.

[0062] Example 2: Synthesis of ethyl 2-((4,4-dimethyl-1-oxobenzothiopyran-6-)ethynyl)-4-hydroxypyrimidine-5-carboxylate (Compound 4)

[0063]

[0064] The compound 3 prepared in Example 1 above was added to a flask, and dry dichloromethane (DCM) was added. m-CPBA (meta-chloroperbenzoic acid) was added under an ice-water bath, and the mixture was stirred and warmed to room temperature. After the reaction was completed as monitored by TLC thin-layer chromatography, DCM was evaporated, and ethyl acetate was added for extraction. The mixture was washed with 2×30 mL of saturated sodium bicarbonate solution and 30 mL of saturated sodium chloride solution, and the organic layer was dried over anhydrous sodium sulfate, filtered and dried to obtain a crude product, which was purified by column chromatography to obtain compound 4.

[0065] The test results of the product prepared above are as follows: 1 H NMR (400 MHz, CDCl3) δ8.06 (d, J = 6.4 Hz, 1H), 7.81 (d, J = 8.1 Hz, 1H), 7.63-7.57 (m, 2H), 7.16 (dd, J = 8.1, 2.1 Hz, 1H), 7.03 (s, 0H), 4.30 (q, J = 6.4 Hz, 2H), 3.18-3.12 (m, 2H), 2.34-2.28 (m, 2H), 1.37 (d, J = 12.7 Hz, 1H), 1.25 (s, 5H); this indicates that compound 4 was synthesized.

[0066] Example 3: Synthesis of ethyl 2-((4,4-dimethyl-1-benzothiopyran-6-)ethynyl)-4-bromopyrimidine-5-carboxylate (Compound 6)

[0067]

[0068] Compound 6-ethynyl-4,4-dimethylthiobenzopyran (i.e., compound 1) and ethyl 2-bromo-4-iodobenzoate (i.e., compound 5) were added to a flask, and Pd(PPh3)2Cl2 and CuI were added. The mixture was protected by argon and the gas was replaced three times to exclude oxygen. 5 ml of dry THF and dried Et3N were added with a syringe, and the reaction was refluxed. After the reaction was completed as monitored by TLC thin-layer chromatography, THF was evaporated, and ethyl acetate was added for extraction. The mixture was washed with 5.0 eq 2N dilute hydrochloric acid, 2×30 mL saturated sodium bicarbonate solution and 30 mL saturated sodium chloride solution, and the organic layer was dried over anhydrous sodium sulfate, filtered and dried to obtain a crude product, which was purified by column chromatography to obtain compound 6.

[0069] The test results of the product prepared above are as follows: 1H NMR (400 MHz, CDCl3) δ7.89 (d, J = 7.9 Hz, 1H), 7.47 (dd, J = 6.2, 2.2 Hz, 1H), 7.36 (d, J = 2.2 Hz, 1H), 7.23 (s, 0H), 7.12-7.07 (m, 2H), 6.69 (s, 2H), 4.30 (q, J = 6.4 Hz, 2H), 3.02-2.96 (m, 2H), 2.07-2.01 (m, 2H), 1.37 (t, J = 6.4 Hz, 3H); this indicated that compound 6 was synthesized.

[0070] Example 4: Synthesis of ethyl 2-((4,4-dimethyl-1-oxobenzothiopyran-6-)ethynyl)-4-bromopyrimidine-5-carboxylate (Compound 7)

[0071]

[0072] The compound 6 prepared in Example 3 above was added to a flask, and dry dichloromethane was added. m-CPBA was added under an ice-water bath, and the mixture was stirred and warmed to room temperature. After the reaction was completed as monitored by TLC thin-layer chromatography, DCM was evaporated, and ethyl acetate was added for extraction. The mixture was washed with 2×30 mL of saturated sodium bicarbonate solution and 30 mL of saturated sodium chloride solution, and the organic layer was dried over anhydrous sodium sulfate, filtered and dried to obtain a crude product, which was purified by column chromatography to obtain compound 7 (yield 85%).

[0073] The test results of the product prepared above are as follows: 1 H NMR (400MHz, CDCl3) δ8.06(d,J=6.4Hz,1H),7.89(d,J=7.9Hz,1H),7.63-7.57(m,2H),7.23(s,0H),7.10(dd,J=7.9,2.2 Hz,1H),6.69(s,2H),4.30(q,J=6.4Hz,2H),3.18-3.12(m,2H),2.34-2.28(m,2H),1.37(d,J=12.7Hz,1H),1.25(s,5H). 13 C NMR (125 MHz, CDCl3) δ 168.47, 150.39, 145.26, 144.99, 132.89, 131.86, 128.63, 128.56, 127.63, 125.81, 119.77, 116.13, 111.80, 89.23, 89.14, 60.25, 49.74, 37.24, 33.34, 30.37, 14.32; this indicated that compound 7 was synthesized.

[0074] Example 5: Synthesis of Intermediate Compound I (Compound 9)

[0075]

[0076] 2.3 g (i.e. 6.4 mmol) of compound 7 and 2.4 g (i.e. 12.8 mmol) of 4-(diethoxymethyl)piperidine (i.e. compound 8) were added into a flask, and the reaction was carried out with methanol as solvent and TsOH (p-toluenesulfonic acid) as catalyst. After the reaction was completed as monitored by TLC thin-layer chromatography, hydrochloric acid was added to react to obtain compound 9.

[0077] The test results of the product prepared above are as follows: 1 H NMR (400MHz, CDCl3) δ9.61(dp,J=7.7,0.9Hz,1H),8.06(d,J=6.4Hz,1H),7.83(d,J=8.3Hz,1H),7. 63-7.57(m,2H),7.30(dd,J=8.2,2.2Hz,1H),7.19(s,0H),4.30(q,J=6.4Hz,2H),3.58(ddd,J=12. 1,8.3,5.6Hz,2H),3.45(ddd,J=11.9,8.3,5.6Hz,2H),3.18-3.12(m,2H),2.46(dp,J=7.7,5.4Hz, 1H),2.34-2.28(m,2H),2.12-2.02(m,2H),1.87-1.77(m,2H),1.37(t,J=6.3Hz,3H),1.25(s,5H). 13 C NMR (125 MHz, CDCl3) δ 205.56, 167.08, 149.90, 145.26, 144.99, 131.86, 130.41, 128.59, 127.72, 127.63, 125.81, 124.25, 122.39, 117.99, 89.39, 89.24, 61.10, 49.74, 48.97, 45.50, 37.24, 33.34, 30.37, 25.55, 14.32; this indicated that compound 9 was synthesized.

[0078] Example 6: Synthesis of Compound MJ-1

[0079]

[0080] 1.2 g (i.e. 1.8 mmol) of compound 9 and 1.2 g (i.e. 2.7 mmol) of compound 10 were dissolved in 30 ml of N,N-dimethylformamide, stirred at room temperature for 1 hour, and then 0.6 g (i.e. 3.6 mmol) of sodium triacetylborohydride was added. The mixture was stirred at room temperature for 8-10 hours. After the reaction was completed as monitored by TLC thin-layer chromatography, saturated ammonium chloride solution was added to quench the reaction. Ethyl acetate and water were added for extraction, and the ethyl acetate phase was washed with saturated brine. After vacuum distillation, separation was performed on a preparative plate to obtain 0.96 g of pure compound MJ-1 with a yield of 48%.

[0081] The test results of the product prepared above are as follows: 1 H NMR(400MHz, CDCl3)δ8.70(s,1H),7.89(s,1H),7.81-7.75(m,2H),7.60(dq,J=6.2,5.2H z,1H),7.49(d,J=9.0Hz,1H),7.09-7.03(m,2H),6.48(ddd,J=8.4,7.7,1.8Hz,1H),6.41( dtd,J=8.4,2.0,1.0Hz,1H),6.11(dt,J=7.5,2.0Hz,1H),5.26(dd,J=4.8,3.2Hz,1H),4.5 4(t,J=9.1Hz,1H),4.27(q,J=6.4Hz,2H),4.10-4.00(m,2H),3.72-3.64(m,1H),3.59-3.5 0(m,2H),3.42(dddd,J=12.8,4.8,3.6,0.8Hz,1H),3.35(tt,J=8.1,1.8Hz,1H),3.22(dd d,J=8.8,5.7,2.9Hz,4H),3.15(p,J=6.5Hz,1H),2.93-2.78(m,2H),2.73-2.47(m,7H),2. 42(d,J=5.3Hz,3H),2.29-1.99(m,6H),1.95-1.82(m,4H),1.78(ddd,J=12.5,7.9,5.3Hz, 1H),1.66-1.39(m,6H),1.42-1.23(m,12H),0.89(d,J=1.4Hz,3H),0.84(d,J=1.6Hz,3H). 13C NMR (125MHz, CDCl3) δ187.22,172.67,171.59,170.54,165.17,163.87,157.51,155.75,153.95,15 1.40,140.94,134.64,132.51,129.31,128.42,123.77,122.95,113.31,109.25,94.16,84.88,66. 20,63.91,61.01,59.05,55.83,54.31,53.13,49.97,48.47,48.16,47.45,46.85,38.58,36.55,34.10,34.03,33.33,32.96,29.89,29.19,26.29,26.10,25.50,24.97,17.21,14.28; This indicates that the synthetic compound MJ-1.

[0082] Example 7: Synthesis of Compound MJ-2

[0083]

[0084] 1.2 g (i.e. 1.8 mmol) of compound 9 and 1.0 g (i.e. 2.7 mmol) of compound 11 were dissolved in 30 ml of N,N-dimethylformamide, stirred at room temperature for 1 hour, and then 0.6 g (i.e. 3.6 mmol) of sodium triacetylborohydride was added. The mixture was stirred at room temperature for 8-10 hours. After the reaction was completed as monitored by TLC thin-layer chromatography, saturated ammonium chloride solution was added to quench the reaction. Ethyl acetate and water were added for extraction, and the ethyl acetate phase was washed with saturated brine. After vacuum distillation, separation was performed on a preparative plate to obtain 0.76 g of pure compound MJ-2 with a yield of 48%.

[0085] The test results of the product prepared above are as follows: 1HNMR(400MHz, CDCl3)δ8.70(s,1H),7.31-7.24(m,2H),7.26-7.19(m,1H),7.23-7.17(m ,3H),6.48(ddd,J=8.4,7.7,1.8Hz,1H),6.41(dtd,J=8.4,2.0,1.0Hz,1H),6.11(dt,J= 7.5,2.0Hz,1H),4.47-4.39(m,1H),4.27(q,J=6.4Hz,2H),4.10-4.00(m,2H),3.95(dd, J=6.0,4.4Hz,1H),3.87(d,J=4.6Hz,1H),3.64-3.50(m,5H),3.47(dd,J=6.2,3.5Hz,2H ),3.39-3.31(m,1H),3.05(dd,J=7.3,6.6Hz,1H),2.93-2.78(m,4H),2.78-2.65(m,2H) ,2.69-2.59(m,1H),2.55(dd,J=6.2,3.5Hz,2H),2.53-2.47(m,2H),2.48(d,J=3.5Hz,1 H),2.03(ddd,J=12.5,7.9,5.4Hz,1H),1.95-1.82(m,4H),1.82-1.64(m,3H),1.63-1.5 4(m,1H),1.37(t,J=6.4Hz,3H),0.89(d,J=1.5Hz,3H),0.88(s,1H),0.90-0.82(m,8H). 13 C NMR(125MHz,Chloroform-d)δ173.94,171.78,165.17,163.87,157.51,155.75,140 .94,137.18,134.64,129.42,128.79,128.42,127.22,123.77,109.25,94.16,84.88 ,70.54,66.20,63.90,61.01,55.77,53.11,51.55,48.47,48.16,46.85,45.98,40.77,37.51,36.55,34.10,32.96,29.19,26.10,24.29,22.30,14.28; This indicates that the synthetic compound MJ-2.

[0086] Example 8: Synthesis of Compound MJ-3

[0087]

[0088] 1.2 g (i.e. 1.8 mmol) of compound 9 and 1.0 g (i.e. 2.7 mmol) of compound 12 were dissolved in 30 ml of N,N-dimethylformamide, stirred at room temperature for 1 hour, and then 0.6 g (i.e. 3.6 mmol) of sodium triacetylborohydride was added. The mixture was stirred at room temperature for 8-10 hours. After the reaction was completed as monitored by TLC thin-layer chromatography, saturated ammonium chloride solution was added to quench the reaction. Ethyl acetate and water were added for extraction, and the ethyl acetate phase was washed with saturated brine. After vacuum distillation, separation was performed on a preparative plate to obtain 0.86 g of pure compound MJ-2 with a yield of 45%.

[0089] The test results of the product prepared above are as follows: 1 H NMR(400MHz, CDCl3)δ8.70(s,1H),7.89(s,1H),7.68-7.56(m,2H),7.52-7.42(m,2H),7.1 3(t,J=2.2Hz,1H),6.73(ddd,J=8.1,2.3,1.3Hz,1H),6.48(ddd,J=8.4,7.7,1.8Hz,1H),6 .41(dtd,J=8.4,2.0,1.0Hz,1H),6.25(t,J=5.7Hz,1H),6.11(dt,J=7.5,2.0Hz,1H),5.26 (dd,J=4.8,3.2Hz,1H),4.54(t,J=9.1Hz,1H),4.27(q,J=6.4Hz,2H),4.08-3.99(m,2H),3 .72-3.64(m,1H),3.60-3.50(m,2H),3.42(dddd,J=12.8,4.8,3.7,0.8Hz,1H),3.35(tt,J =8.1,1.8Hz,1H),3.19(d,J=4.4Hz,1H),3.17(d,J=4.5Hz,1H),3.14(q,J=6.5Hz,1H),2.9 3-2.78(m,2H),2.73-2.65(m,1H),2.42(d,J=5.3Hz,3H),2.29-1.99(m,6H),1.98-1.73(m ,7H),1.66-1.39(m,6H),1.43-1.23(m,12H),0.89(d,J=1.4Hz,3H),0.84(d,J=1.6Hz,3H). 13C NMR(125MHz,Chloroform-d)δ186.74,172.67,171.59,170.54,165.17,163.87,157.51,155.75,151. 08,148.68,140.94,135.37,134.64,131.01,128.42,124.83,123.77,122.94,117.88,115.75,109.25 ,94.16,84.88,66.20,61.01,59.05,55.83,54.31,48.47,48.16,48.10,47.45,46.82,38.58,36.55,34.33,34.10,34.03,33.33,29.89,28.93,26.29,26.10,25.50,24.97,17.21,14.28; This indicates that the synthetic compound MJ-3.

[0090] Example 9: Synthesis of Compound MJ-4

[0091]

[0092] 1.2 g (i.e. 1.8 mmol) of compound 9 and 0.6 g (i.e. 2.7 mmol) of compound 13 were dissolved in 30 ml of N,N-dimethylformamide, and stirred at room temperature for 1 hour. Then, 0.6 g (i.e. 3.6 mmol) of sodium triacetylborohydride was added and stirred at room temperature for 8-10 hours. After the reaction was completed as monitored by TLC thin-layer chromatography, saturated ammonium chloride solution was added to quench the reaction. After extraction with ethyl acetate and water, the ethyl acetate phase was washed with saturated brine, and the mixture was separated by preparative plate after vacuum distillation to obtain 0.56 g of pure compound MJ-4 with a yield of 33%.

[0093] The test results of the product prepared above are as follows: 1H NMR (400MHz, Chloroform-d) δ8.70 (s, 1H), 7.33-7.17 (m, 6H), 6.73 (t, J = 5.9Hz, 1H), 6.48 (ddd, J = 8.4, 7.7, 1.8Hz, 1H), 6.41 (dtd, J = 8.4, 2.0, 1. 0Hz,1H),6.11(dt,J=7.5,2.0Hz,1H),4.35-4.26(m,2H),4.26(d,J=6.4Hz,1H),4.04(ddd,J=12.5,8.9,6.3Hz,2H),3.97(dd,J=6.1,4.5Hz,1H), 3.87(d,J=4.5Hz,1H),3.58(ddd,J=12.5,8.8,6.2Hz,3H),3.39-3.31(m,1H),3.11(dd,J=5.9,4.9Hz,2H),3.05(dd,J=7.3,6.6Hz,1H),2.93-2.7 8(m,4H),2.78-2.65(m,2H),2.03(ddd,J=12.5,7.9,5.4Hz,1H),1.94-1. 71(m,7H),1.65-1.51(m,2H),1.37(t,J=6.4Hz,3H),0.91-0.82(m,12H). 13 C NMR(125MHz,Chloroform-d)δ173.89,173.67,165.17,163.87,157.51,155.75, 140.94,137.18,134.64,129.42,128.79,128.42,127.22,123.77,109.25,94.16 ,84.88,70.54,66.20,61.01,55.77,52.62,48.47,48.16,46.82,44.88,40.73,37.51,36.55,34.77,34.10,28.74,26.10,24.99,22.40,14.28; This indicates that the synthetic compound MJ-4.

[0094] Example 10: Synthesis of Compound MJ-5

[0095]

[0096] 1.2 g (i.e. 1.8 mmol) of compound 9 and 0.8 g (i.e. 2.7 mmol) of compound 14 were dissolved in 30 ml of N,N-dimethylformamide, stirred at room temperature for 1 hour, and then 0.6 g (i.e. 3.6 mmol) of sodium triacetylborohydride was added. The mixture was stirred at room temperature for 8-10 hours. After the reaction was completed as monitored by TLC thin-layer chromatography, a saturated ammonium chloride solution was added to quench the reaction. After extraction with ethyl acetate and water, the ethyl acetate phase was washed with saturated brine, and the mixture was separated by preparative plate after vacuum distillation to obtain 0.75 g of pure compound MJ-5 with a yield of 36%.

[0097] The test results of the product prepared above are as follows: 1 H NMR(400MHz,Chloroform-d)δ8.70(s,1H),7.89(s,1H),7.81-7.75(m,2H),7.60(dq,J=6.2,5.2Hz,1H),7.4 9(d,J=9.0Hz,1H),7.09-7.03(m,2H),6.48(ddd,J=8.4,7.7,1.8Hz,1H),6.41(dtd,J=8.4,2.0,1.0Hz,1H), 6.11(dt,J=7.5,2.0Hz,1H),5.26(dd,J=4.8,3.2Hz,1H),4.54(t,J=9.1Hz,1H),4.27(q,J=6.4Hz,2H),4.10 -4.00(m,2H),3.72-3.64(m,1H),3.59-3.50(m,4H),3.50(dd,J=5.4,2.7Hz,2H),3.42(dddd,J=12.8,4.8,3. 6,0.8Hz,1H),3.35(tt,J=8.1,1.8Hz,1H),3.15(p,J=6.5Hz,1H),2.88(ddd,J=12.3,7.9,5.4Hz,1H),2.83( ddd,J=12.5,7.9,5.4Hz,1H),2.79(s,1H),2.76(s,1H),2.72-2.63(m,4H),2.57-2.51(m,1H),2.42(d,J=5. 3Hz,3H),2.29-2.20(m,1H),2.20-2.03(m,4H),1.99-1.81(m,7H),1.81-1.73(m,1H),1.74(d,J=2.6Hz,1H) ,1.73(d,J=2.7Hz,1H),1.66-1.39(m,6H),1.43-1.23(m,12H),0.89(d,J=1.4Hz,3H),0.84(d,J=1.6Hz,3H). 13C NMR(125MHz,Chloroform-d)δ187.22,172.67,171.59,170.54,165.17,163.87,157.51,155.75,152.96, 151.40,140.94,134.64,132.50,129.31,128.42,123.77,122.95,113.30,109.25,94.16,84.88,66.20, This indicates that the synthetic compound MJ-5.

[0098] Example 11: Synthesis of Compound MJ-6

[0099]

[0100] 1.2 g (i.e. 1.8 mmol) of compound 9 and 0.8 g (i.e. 2.7 mmol) of compound 15 were dissolved in 30 ml of N,N-dimethylformamide, and stirred at room temperature for 1 hour. Then, 0.6 g (i.e. 3.6 mmol) of sodium triacetylborohydride was added, and the mixture was stirred at room temperature for 8-10 hours. After the reaction was completed as monitored by TLC thin-layer chromatography, saturated ammonium chloride solution was added to quench the reaction. After extraction with ethyl acetate and water, the ethyl acetate phase was washed with saturated brine, and the mixture was separated by preparative plate after vacuum distillation to obtain 0.84 g of pure compound MJ-6 with a yield of 38%.

[0101] The test results of the product prepared above are as follows: 1H NMR(400MHz,Chloroform-d)δ8.70(s,1H),7.31-7.24(m,2H),7.26-7.17(m,4H ),6.48(ddd,J=8.4,7.7,1.8Hz,1H),6.41(dtd,J=8.4,2.0,1.0Hz,1H),6.11(dt ,J=7.5,2.0Hz,1H),4.47-4.39(m,1H),4.27(q,J=6.4Hz,2H),4.09-4.00(m,2H ),3.95(dd,J=6.0,4.4Hz,1H),3.87(d,J=4.6Hz,1H),3.59(dt,J=12.7,6.3Hz,1 H),3.58-3.52(m,1H),3.55-3.50(m,1H),3.47(ddd,J=12.5,5.7,3.1Hz,2H),3 .41-3.31(m,3H),3.05(dd,J=7.3,6.6Hz,1H),2.93-2.63(m,11H),2.57-2.51(m ,1H),2.03(ddd,J=12.5,7.9,5.4Hz,1H),1.95-1.64(m,12H),1.63-1.54(m,1H ),1.37(t,J=6.4Hz,3H),0.89(d,J=1.5Hz,3H),0.88(s,1H),0.90-0.82(m,8H). 13 C NMR(125MHz,Chloroform-d)δ173.94,171.90,165.17,163.87,157.51,155.75,140.94 ,137.18,134.64,129.42,128.79,128.42,127.22,123.77,109.25,94.16,84.88,70.54 ,66.20,63.93,61.01,59.97,55.77,51.55,48.47,48.16,46.85,42.73,40.77,37.51,36.55,34.38,34.10,31.93,31.06,29.19,26.10,24.29,22.30,14.28; This indicates that the synthetic compound MJ-6.

[0102] Example 12: Synthesis of Compound MJ-7

[0103]

[0104] 1.2 g (i.e. 1.8 mmol) of compound 9 and 0.9 g (i.e. 2.7 mmol) of compound 16 were dissolved in 30 ml of N,N-dimethylformamide, and stirred at room temperature for 1 hour. Then, 0.6 g (i.e. 3.6 mmol) of sodium triacetylborohydride was added, and the mixture was stirred at room temperature for 8-10 hours. After the reaction was completed as monitored by TLC thin-layer chromatography, saturated ammonium chloride solution was added to quench the reaction. After extraction with ethyl acetate and water, the ethyl acetate phase was washed with saturated brine, and the mixture was separated by preparative plate after vacuum distillation to obtain 0.63 g of pure compound MJ-7 with a yield of 33%.

[0105] The test results of the product prepared above are as follows: 1 H NMR(400MHz,Chloroform-d)δ8.70(s,1H),7.89(s,1H),7.81-7.75(m,2H),7.60(dq,J=6.2,5.2Hz,1H ),7.49(d,J=9.0Hz,1H),7.09-7.03(m,2H),6.48(ddd,J=8.4,7.7,1.8Hz,1H),6.41(dtd,J=8.4,2.0,1 .0Hz,1H),6.11(dt,J=7.5,2.0Hz,1H),5.26(dd,J=4.8,3.2Hz,1H),4.54(t,J=9.1Hz,1H),4.27(q,J=6 .4Hz,2H),4.10-4.00(m,2H),3.72-3.64(m,1H),3.59-3.38(m,7H),3.35(tt,J=8.1,1.8Hz,1H),3.15( p,J=6.5Hz,1H),2.93-2.78(m,2H),2.73-2.65(m,2H),2.65(d,J=3.7Hz,1H),2.61(dd,J=11.4,4.6Hz ,1H),2.56-2.50(m,1H),2.47(dd,J=6.5,3.7Hz,2H),2.42(d,J=5.3Hz,3H),2.29-2.20(m,1H),2.20-2 .03(m,4H),1.99-1.72(m,9H),1.69(dd,J=6.1,3.4Hz,2H),1.63(d,J=3.7Hz,1H),1.64-1.56(m,3H),1 .59-1.55(m,2H),1.55-1.45(m,3H),1.48-1.23(m,13H),0.89(d,J=1.4Hz,3H),0.84(d,J=1.6Hz,3H). 13C NMR(125MHz,Chloroform-d)δ187.22,172.67,171.59,170.54,165.17,163.87,157.51,155.75,152.83,1 51.40,140.94,134.64,132.50,129.31,128.42,123.77,122.95,113.30,109.25,94.16,84.88,66.20,63. This indicates that the synthetic compound MJ-7 is obtained.

[0106] Example 13: Synthesis of Compound MJ-8

[0107]

[0108] 1.2 g (i.e. 1.8 mmol) of compound 9 and 0.9 g (i.e. 2.7 mmol) of compound 17 were dissolved in 30 ml of N,N-dimethylformamide, stirred at room temperature for 1 hour, and then 0.6 g (i.e. 3.6 mmol) of sodium triacetylborohydride was added. The mixture was stirred at room temperature for 8-10 hours. After the reaction was completed as monitored by TLC thin-layer chromatography, saturated ammonium chloride solution was added to quench the reaction, and ethyl acetate and water were added for extraction. The ethyl acetate phase was washed with saturated brine, and the mixture was separated by preparative plate after vacuum distillation to obtain 0.58 g of pure compound MJ-8 with a yield of 30%.

[0109] The test results of the product prepared above are as follows: 1H NMR(400MHz,Chloroform-d)δ8.70(s,1H),7.41(d,J=9.3Hz,1H),7.31-7.17(m,5H),6.48(ddd,J=8 .4,7.7,1.8Hz,1H),6.41(dtd,J=8.4,2.0,1.0Hz,1H),6.11(dt,J=7.5,2.0Hz,1H),4.35-4.26(m,2 H),4.26(d,J=6.4Hz,1H),4.10-4.00(m,2H),3.95(dd,J=6.0,4.4Hz,1H),3.87(d,J=4.6Hz,1H),3. 64-3.43(m,5H),3.42-3.35(m,2H),3.38-3.31(m,1H),3.05(dd,J=7.3,6.6Hz,1H),2.93-2.78(m,4H ),2.78-2.70(m,1H),2.74-2.66(m,1H),2.67(d,J=3.7Hz,1H),2.65(d,J=3.7Hz,1H),2.61(dd,J=1 1.4,4.6Hz,1H),2.56-2.50(m,1H),2.47(dd,J=6.5,3.8Hz,2H),2.03(ddd,J=12.5,7.9,5.4Hz,1H), 1.95-1.70(m,8H),1.69(dd,J=13.7,6.6Hz,1H),1.62(ddd,J=6.6,3.7,0.9Hz,4H),1.56(dd,J=6.6 ,3.7Hz,2H),1.41-1.30(m,5H),0.94(t,J=7.3Hz,3H),0.89(d,J=1.5Hz,3H),0.84(d,J=1.6Hz,3H). 13 C NMR(125MHz,Chloroform-d)δ173.90,172.38,165.17,163.87,157.51,155.75,140.94,1 37.18,134.64,129.42,128.79,128.42,127.22,123.77,109.25,94.16,84.88,71.30,66 .20,63.93,61.01,55.77,52.71,50.71,48.47,48.16,46.85,42.81,37.51,36.55,36.23,35.46,34.10,33.85,33.40,32.97,29.19,26.10,18.85,14.28,13.44; This indicates that the synthetic compound MJ-8.

[0110] Example 14: Synthesis of ethyl 2-((4,4-dimethyl-1-oxobenzothiopyran-6-)ethynyl)-4-hydroxypyrimidine-5-carboxylate (Compound 19)

[0111]

[0112] 1.2 g (i.e. 1.2 mmol) of compound 4 and 0.7 g (i.e. 2.4 mmol) of compound 20 were dissolved in 30 ml of N,N-dimethylformamide, and K2CO3 was added and stirred at 90°C. After TLC thin-layer chromatography monitored the reaction to be complete, saturated ammonium chloride solution was added to quench the reaction, ethyl acetate and water were added for extraction, the ethyl acetate phase was washed with saturated brine, and dichloromethane was added to dissolve after reduced pressure distillation, 1M HCl was added to remove the protecting group, and the mixture was stirred at room temperature for 8-10 hours. After TLC thin-layer chromatography monitored the reaction to be complete, dichloromethane was removed by rotary distillation, ethyl acetate and water were added for extraction, the ethyl acetate phase was washed with saturated brine, and the compound 19 was separated by preparative plate after reduced pressure distillation to obtain 1.1 g of pure compound 19 with a yield of 68%.

[0113] The test results of the product prepared above are as follows: 1 HNMR(400MHz,Chloroform-d)δ8.06(d,J=6.4Hz,1H),7.86(d,J=8.0Hz,1H),7.63-7.57(m,2H),7.23(dd,J=8.0,2.1Hz,1H),7.10(s,0H),4. 30(q,J=6.4Hz,2H),4.16(t,J=4.0Hz,2H),3.21-3.12(m,4H),2.34-2.28(m,2H),1.73(d,J=13.7Hz,1H),1.37(t,J=6.3Hz,3H),1.25(s,5H). 13 C NMR (125 MHz, Chloroform-d) δ 166.09, 159.34, 145.26, 144.99, 131.86, 131.39, 128.62, 127.63, 127.35, 125.82, 125.57, 120.84, 114.53, 89.25, 89.20, 70.94, 61.11, 49.74, 41.81, 37.24, 33.34, 30.37, 14.32; this indicated that compound 19 was synthesized.

[0114] Example 15: Synthesis of ethyl 2-((4,4-dimethyl-1-oxobenzothiopyran-6-)ethynyl)-4-hydroxypyrimidine-5-carboxylate (Compound 21)

[0115]

[0116] 1.2 g (i.e. 1.2 mmol) of compound 4 and 0.8 g (i.e. 2.4 mmol) of compound 20 were dissolved in 30 ml of N,N-dimethylformamide, and K2CO3 was added and stirred at 90°C. After TLC thin-layer chromatography monitored the reaction to be complete, saturated ammonium chloride solution was added to quench the reaction, ethyl acetate and water were added for extraction, the ethyl acetate phase was washed with saturated brine, and dichloromethane was added to dissolve after reduced pressure distillation, 1M HCl was added to remove the protecting group, and the mixture was stirred at room temperature for 8-10 hours. After TLC thin-layer chromatography monitored the reaction to be complete, dichloromethane was removed by rotary distillation, ethyl acetate and water were added for extraction, the ethyl acetate phase was washed with saturated brine, and the compound 21 was separated by preparative plate after reduced pressure distillation to obtain 1.1 g of pure compound 21 with a yield of 75%.

[0117] The test results of the product prepared above are as follows: 1 HNMR(500MHz,Chloroform-d)δ8.06(d,J=6.4Hz,1H),7.86(d,J=8.0Hz,1H),7.63- 7.57(m,2H),7.23(dd,J=8.0,2.1Hz,1H),7.11(s,0H),4.30(q,J=6.4Hz,2H),4.05( t,J=5.7Hz,2H),3.18-3.12(m,2H),2.85(tt,J=6.3,5.6Hz,2H),2.34-2.28(m,2H) ,2.16(d,J=12.8Hz,1H),1.93(p,J=5.6Hz,2H),1.37(t,J=6.3Hz,3H),1.25(s,5H). 13 C NMR (125 MHz, Chloroform-d) δ 166.26, 159.65, 145.26, 144.99, 131.86, 131.45, 128.62, 127.63, 127.38, 125.82, 125.57, 121.02, 114.36, 89.25, 89.20, 67.70, 61.11, 49.74, 37.24, 36.42, 33.34, 32.70, 30.37, 14.32; this indicated that compound 21 was synthesized.

[0118] Example 16: Synthesis of Compound MJ-9

[0119]

[0120] 1.1 g (i.e. 1.2 mmol) of compound 19 and 0.9 g (i.e. 1.3 mmol) of compound 22 were dissolved in 30 ml of N,N-dimethylformamide, and stirred at room temperature for 1 hour. Then, 0.6 g (i.e. 2.4 mmol) of HATU (N,N,N′,N′-tetramethyl-O-(7-azabenzotriazole-1-yl)urea hexafluorophosphate) was added, and stirred at room temperature for 8-10 hours. After the reaction was completed as monitored by TLC thin-layer chromatography, saturated ammonium chloride solution was added to quench the reaction. Ethyl acetate and water were added for extraction, and the ethyl acetate phase was washed with saturated brine. After vacuum distillation, separation was performed on a preparative plate to obtain 0.8 g of pure compound MJ-9 with a yield of 43%.

[0121] The test results of the product prepared above are as follows: 1 HNMR(400MHz,Chloroform-d)δ8.99(s,1H),7.45(t,J=4.8Hz,1H),7.33-7.17(m,6H),7.03(t,J=4.9Hz,1H),6.48(ddd,J=8.4,7.7, 1.8Hz,1H),6.41(dtd,J=8.4,2.0,1.0Hz,1H),6.11(dt,J=7.5,2.0Hz,1H),4.66(d,J=1.3Hz,2H),4.36-4.25(m,3H),3.95(dd,J=6. 0,4.4Hz,1H),3.87(d,J=4.6Hz,1H),3.59(h,J=6.3Hz,1H),3.40-3.25(m,5H),3.05(dd,J=7.3,6.6Hz,1H),2.93-2.78(m,4H),2.78 -2.65(m,2H),2.03(ddd,J=12.5,7.9,5.4Hz,1H),1.87-1.73(m,2H),1.66-1.51(m,2H),1.38(t,J=6.3Hz,3H),0.91-0.82(m,11H). 13 C NMR(125MHz,Chloroform-d)δ173.89,173.82,169.69,165.10,163.40,157.21, 156.74,140.80,137.18,134.65,129.42,128.79,128.42,127.22,123.80,109.7 3,94.22,84.89,70.54,67.37,66.20,61.01,55.77,52.57,48.47,48.16,40.73,39.52,39.07,37.51,36.55,34.10,26.10,24.99,22.40,14.28; This indicates that the synthetic compound MJ-9.

[0122] Example 17: Synthesis of Compound MJ-10

[0123]

[0124] 1.1 g (i.e. 1.2 mmol) of compound 19 and 1.4 g (i.e. 1.3 mmol) of compound 22 were dissolved in 30 ml of N,N-dimethylformamide, stirred at room temperature for 1 hour, and then 0.6 g (i.e. 2.4 mmol) of HATU was added. The mixture was stirred at room temperature for 8-10 hours. After the reaction was completed as monitored by TLC thin-layer chromatography, saturated ammonium chloride solution was added to quench the reaction. Ethyl acetate and water were added for extraction, and the ethyl acetate phase was washed with saturated brine. After vacuum distillation, separation was performed on a preparative plate to obtain 0.86 g of pure compound MJ-10 with a yield of 42%.

[0125] The test results of the product prepared above are as follows: 1HNMR(400MHz,Chloroform-d)δ8.99(s,1H),8.20(t,J=2.2Hz,1H),7.94(ddd,J=8.2,2.2,1.1Hz,1H),7.91(ddd,J=7.9,2.2,1.2Hz,1H),7.90-7.84(m,2H),7.67(t,J=8.0Hz,1H),7.60(dq,J=6.2,5.2Hz,1H),7.52-7.44(m,2H),6.48(ddd,J=8.4,7.7,1.8Hz,1H),6.41(dtd,J=8.4,2.0,1.0Hz,1H),6.11(dt,J=7.5,2.0Hz,1H),5.26(dd,J=4.8,3.2Hz,1H),4.66(d,J=1.3Hz,2H),4.54(t,J=9.1Hz,1H),4.32(q,J=6.3Hz,2H),3.72-3.64(m,1H),3.53(dtd,J=5.3,4.3,2.2Hz,2H),3.46-3.38(m,3H),3.35(tt,J=8.1,1.8Hz,1H),3.15(p,J=6.5Hz,1H),2.93-2.78(m,2H),2.73-2.65(m,1H),2.42(d,J=5.3Hz,3H),2.29-2.20(m,1H),2.20-2.03(m,4H),1.95(ddd,J=12.5,7.8,5.4Hz,1H),1.77(ddd,J=12.5,7.9,5.3Hz,1H),1.59(dddd,J=11.2,8.6,5.7,4.0Hz,2H),1.55-1.47(m,2H),1.51-1.46(m,1H),1.49-1.39(m,1H),1.39(s,1H),1.38(s,2H),1.38-1.31(m,3H),1.34-1.23(m,6H),0.89(d,J=1.4Hz,3H),0.84(d,J=1.6Hz,3H). 13CNMR(125MHz,Chloroform-d)δ187.05,172.67,171.59,170.54,169.69,167.85,165.10,163.40,157.21 ,156.74,151.55,140.80,135.49,134.65,134.23,133.57,131.03,130.73,129.85,128.42,123.80,122 .94,109.73,94.22,84.89,67.37,66.20,61.01,59.05,55.83,54.31,48.47,48.16,47.45,39.46,39.25,38.58,36.55,34.10,34.03,33.33,29.89,26.29,26.10,25.50,24.97,17.21,14.28; This indicates that the synthetic compound MJ-10.

[0126] Example 18: Synthesis of Compound MJ-11

[0127]

[0128] 1.1 g (i.e. 1.2 mmol) of compound 19 and 0.9 g (i.e. 1.3 mmol) of compound 23 were dissolved in 30 ml of N,N-dimethylformamide, and stirred at room temperature for 1 hour. Then, 0.6 g (i.e. 3.6 mmol) of HATU was added and stirred at room temperature for 8-10 hours. After the reaction was completed as monitored by TLC thin-layer chromatography, saturated ammonium chloride solution was added to quench the reaction. After extraction with ethyl acetate and water, the ethyl acetate phase was washed with saturated brine, and the mixture was separated by preparative plate after vacuum distillation to obtain 0.66 g of pure compound MJ-11 with a yield of 32.0%.

[0129] The test results of the product prepared above are as follows: 1H NMR(400MHz,Chloroform-d)δ8.99(s,1H),7.33-7.17(m,6H),6.92(t,J=4.8Hz,1H),6.82(t,J=4.9Hz,1H),6.48(ddd,J=8.4,7.7,1.8Hz,1H),6.4 1(dtd,J=8.4,2.0,1.0Hz,1H),6.11(dt,J=7.5,2.0Hz,1H),4.66(d,J=1. 3Hz,2H),4.33(d,J=6.3Hz,1H),4.33-4.25(m,2H),3.95(dd,J=6.0,4.4Hz ,1H),3.87(d,J=4.6Hz,1H),3.59(h,J=6.3Hz,1H),3.39-3.31(m,1H),3. 24-3.06(m,4H),3.05(dd,J=7.3,6.6Hz,1H),2.93-2.78(m,4H),2.78-2. 65(m,2H),2.03(ddd,J=12.5,7.9,5.4Hz,1H),1.87-1.73(m,2H),1.65-1 .51(m,2H),1.50-1.41(m,4H),1.38(t,J=6.3Hz,3H),0.91-0.82(m,12H). 13 C NMR(125MHz,Chloroform-d)δ173.89,172.55,169.83,165.10,163.40,157.21,156. 74,140.80,137.18,134.65,129.42,128.79,128.42,127.22,123.80,109.73,94.22 ,84.89,70.54,67.37,66.20,61.01,55.77,52.57,48.47,48.16,40.73,40.12,39.66,37.51,36.55,34.10,26.71,26.61,26.10,24.99,22.40,14.28; This indicates that the synthetic compound MJ-11.

[0130] Example 19: Synthesis of Compound MJ-12

[0131]

[0132] 1.1 g (i.e. 1.2 mmol) of compound 19 and 1.0 g (i.e. 1.2 mmol) of compound 22 were dissolved in 30 ml of N,N-dimethylformamide, stirred at room temperature for 1 hour, and then 0.6 g (i.e. 2.4 mmol) of HATU was added. The mixture was stirred at room temperature for 8-10 hours. After the reaction was completed as monitored by TLC thin-layer chromatography, saturated ammonium chloride solution was added to quench the reaction. Ethyl acetate and water were added for extraction, and the ethyl acetate phase was washed with saturated brine. After vacuum distillation, separation was performed on a preparative plate to obtain 0.85 g of pure compound MJ-12 with a yield of 45%.

[0133] The test results of the product prepared above are as follows: 1 HNMR(400MHz,Chloroform-d)δ8.99(s,1H),8.20(t,J=2.2Hz,1H),7.92(dddd,J=15.4,7.9,2.2,1.2Hz,2H ),7.89(s,1H),7.73(t,J=5.0Hz,1H),7.67(t,J=8.0Hz,1H),7.60(dq,J=6.2,5.2Hz,1H),7.49(d,J=9.0Hz ,1H),6.92(t,J=4.8Hz,1H),6.48(ddd,J=8.4,7.7,1.8Hz,1H),6.41(dtd,J=8.4,2.0,1.0Hz,1H),6.11(dt ,J=7.5,2.0Hz,1H),5.26(dd,J=4.8,3.2Hz,1H),4.66(d,J=1.3Hz,2H),4.54(t,J=9.1Hz,1H),4.32(q,J=6. 3Hz,2H),3.72-3.64(m,1H),3.46-3.37(m,1H),3.40-3.29(m,3H),3.19(qd,J=5.1,2.4Hz,2H),3.18-3.10 (m,1H),2.93-2.78(m,2H),2.73-2.65(m,1H),2.42(d,J=5.3Hz,3H),2.29-2.20(m,1H),2.20-2.03(m,4H), 1.95(ddd,J=12.5,7.8,5.4Hz,1H),1.77(ddd,J=12.5,7.9,5.3Hz,1H),1.66-1.58(m,1H),1.61-1.57(m,2 H),1.58-1.54(m,1H),1.57-1.40(m,6H),1.43-1.23(m,12H),0.89(d,J=1.4Hz,3H),0.84(d,J=1.6Hz,3H). 13C NMR(125MHz,Chloroform-d)δ187.05,172.67,171.59,170.54,169.83,167.97,165.10,163.40,157.21,15 6.74,151.55,140.80,135.48,134.65,134.22,133.57,131.03,130.75,129.85,128.42,123.80,122.94,10 This indicates that the synthetic compound MJ-12 is obtained.

[0134] Example 20: Synthesis of Compound MJ-13

[0135]

[0136] 1.1 g (i.e. 1.2 mmol) of compound 19 and 0.9 g (i.e. 1.2 mmol) of compound 25 were dissolved in 30 ml of N,N-dimethylformamide, and stirred at room temperature for 1 hour. Then, 0.6 g (i.e. 2.4 mmol) of HATU was added and stirred at room temperature for 8-10 hours. After the reaction was completed as monitored by TLC thin-layer chromatography, saturated ammonium chloride solution was added to quench the reaction. After extraction with ethyl acetate and water, the ethyl acetate phase was washed with saturated brine, and the mixture was separated by preparative plate after vacuum distillation to obtain 0.72 g of pure compound MJ-13 with a yield of 39.0%.

[0137] The test results of the product prepared above are as follows: 1H NMR(400MHz,Chloroform-d)δ8.99(s,1H),7.33-7.17(m,6H),6.92(t,J=4.8Hz,1H),6.82(t,J=4.9Hz,1H),6.48(ddd,J=8.4,7.7,1.8Hz,1H), 6.41(dtd,J=8.4,2.0,1.0Hz,1H),6.11(dt,J=7.5,2.0Hz,1H),4.66(d,J=1.3Hz,2H),4.33(d,J=6.3Hz,1H),4.33-4.25(m,2H),3.97(dd,J=6.1 ,4.5Hz,1H),3.87(d,J=4.5Hz,1H),3.59(h,J=6.3Hz,1H),3.39-3.31(m ,1H),3.24-3.02(m,5H),2.93-2.78(m,4H),2.78-2.65(m,2H),2.03(dd d,J=12.5,7.9,5.4Hz,1H),1.87-1.73(m,2H),1.66-1.51(m,2H),1.54- 1.48(m,1H),1.51-1.42(m,3H),1.41-1.29(m,7H),0.91-0.82(m,11H). 13 CNMR(125MHz,Chloroform-d)δ173.89,172.55,169.83,165.10,163.40,157.21,156.74 ,140.80,137.18,134.65,129.42,128.79,128.42,127.22,123.80,109.73,94.22,84.8 9,70.54,67.37,66.20,61.01,55.77,52.57,48.47,48.16,40.73,40.29,39.84,37.51,36.55,34.10,31.38,28.94,26.54,26.52,26.10,24.99,22.40,14.28; This indicates that the synthetic compound MJ-13.

[0138] Example 21: Synthesis of Compound MJ-14

[0139]

[0140] 1.1 g (i.e. 1.2 mmol) of compound 19 and 0.9 g (i.e. 1.3 mmol) of compound 26 were dissolved in 30 ml of N,N-dimethylformamide, and stirred at room temperature for 1 hour. Then, 0.6 g (i.e. 2.4 mmol) of HATU was added and stirred at room temperature for 8-10 hours. After the reaction was completed as monitored by TLC thin-layer chromatography, saturated ammonium chloride solution was added to quench the reaction. After extraction with ethyl acetate and water, the ethyl acetate phase was washed with saturated brine, and the mixture was separated by preparative plate after reduced pressure distillation to obtain 0.72 g of pure compound MJ-14 with a yield of 43%.

[0141] The test results of the product prepared above are as follows: 1 HNMR(400MHz,Chloroform-d)δ8.99(s,1H),8.20(t,J=2.2Hz,1H),7.92(dddd,J=15.4,7.9,2.2,1.2Hz ,2H),7.89(s,1H),7.74(t,J=5.0Hz,1H),7.67(t,J=8.0Hz,1H),7.60(dq,J=6.2,5.2Hz,1H),7.49(d,J= 9.0Hz,1H),6.92(t,J=4.8Hz,1H),6.48(ddd,J=8.4,7.7,1.8Hz,1H),6.41(dtd,J=8.4,2.0,1.0Hz,1H) ,5.94(dt,J=7.6,2.0Hz,1H),5.26(dd,J=4.8,3.2Hz,1H),4.66(d,J=1.3Hz,2H),4.54(s,0H),4.32(q,J =6.3Hz,2H),3.72-3.64(m,1H),3.42(dddd,J=12.1,4.1,3.3,1.0Hz,1H),3.42-3.31(m,1H),3.30(qd, J=5.4,1.2Hz,2H),3.19(tdd,J=5.7,4.8,2.5Hz,2H),3.19-3.10(m,1H),2.93-2.78(m,2H),2.73-2.65( m,1H),2.42(d,J=5.3Hz,3H),2.29-2.03(m,5H),1.95(ddd,J=12.5,7.8,5.4Hz,1H),1.77(ddd,J=12.5 ,7.9,5.3Hz,1H),1.66-1.40(m,10H),1.43-1.23(m,16H),0.89(d,J=1.5Hz,3H),0.84(d,J=1.7Hz,3H). 13CNMR(125MHz,Chloroform-d)δ187.05,172.67,171.59,170.54,169.83,167.96,165.10,163.40,157.21,156.7 4,151.55,140.87,135.48,134.65,134.21,133.57,131.03,130.75,129.85,128.42,123.80,122.94,109.73,9 This indicates that the synthetic compound MJ-14 is obtained.

[0142] Example 22: Synthesis of Compound MJ-15

[0143]

[0144] 1.1 g (i.e. 1.2 mmol) of compound 19 and 1.2 g (i.e. 1.3 mmol) of compound 27 were dissolved in 30 ml of N,N-dimethylformamide and stirred at room temperature for 1 hour. Then, 0.6 g (i.e. 2.4 mmol) of HATU was added and stirred at room temperature for 8-10 hours. After the reaction was completed as monitored by TLC thin-layer chromatography, saturated ammonium chloride solution was added to quench the reaction. After extraction with ethyl acetate and water, the ethyl acetate phase was washed with saturated brine. After vacuum distillation and separation on a preparative plate, 0.54 g of pure compound MJ-15 was obtained with a yield of 36%.

[0145] The test results of the product prepared above are as follows: 1H NMR(400MHz,Chloroform-d)δ8.98(s,1H),7.33-7.14(m,7H),6.91(t,J=4.9Hz,1H),6.48(ddd,J=8.4,7.7,1.8Hz,1H),6.41(dtd,J=8.4,2 .0,1.0Hz,1H),6.11(dt,J=7.5,2.0Hz,1H),4.36-4.23(m,5H),4.05(s,1H),3.95(dd,J=6.0,4.4Hz,1H),3.87(d,J=4.6Hz,1H),3.78-3.67( m,2H),3.67-3.51(m,9H),3.44-3.30(m,3H),3.24(td,J=5.9,4.8Hz,2H),3.05(dd,J=7.3,6.6Hz,1H),2.93-2.85(m,1H),2.88-2.78(m,3H) ,2.78-2.65(m,2H),2.11-2.04(m,2H),2.07-1.99(m,1H),1.87-1.73 (m,2H),1.66-1.51(m,2H),1.38(t,J=6.3Hz,3H),0.91-0.82(m,11H). 13 C NMR(125MHz,Chloroform-d)δ173.89,173.87,171.75,165.09,164.08,156.92,156.71,140. 80,137.18,134.65,129.42,128.79,128.42,127.22,123.80,110.16,94.22,84.89,70.59,70 .54,70.30,69.67,69.61,69.57,69.48,67.03,66.20,61.01,55.77,52.57,48.47,48.16,40.73,40.36,37.51,37.43,36.55,34.10,29.01,26.10,24.99,22.40,14.28; This indicates that the synthetic compound MJ-15.

[0146] Example 21: Synthesis of Compound MJ-16

[0147]

[0148] 1.2 g (i.e. 1.2 mmol) of compound 19 and 1.3 g (i.e. 1.3 mmol) of compound 22 were dissolved in 30 ml of N,N-dimethylformamide, stirred at room temperature for 1 hour, and then 0.6 g (i.e. 2.4 mmol) of HATU was added. The mixture was stirred at room temperature for 8-10 hours. After the reaction was completed as monitored by TLC thin-layer chromatography, saturated ammonium chloride solution was added to quench the reaction. Ethyl acetate and water were added for extraction, and the ethyl acetate phase was washed with saturated brine. After vacuum distillation, separation was performed on a preparative plate to obtain 0.52 g of pure compound MJ-16 with a yield of 34%.

[0149] The test results of the product prepared above are as follows: 1 HNMR(400MHz,Chloroform-d)δ8.98(s,1H),8.20(t,J=2.2Hz,1H),7.97-7.83(m,4H),7.67(t,J=8.0Hz,1H) ,7.60(dq,J=6.2,5.2Hz,1H),7.49(d,J=9.0Hz,1H),6.91(t,J=4.9Hz,1H),6.48(ddd,J=8.4,7.7,1.8Hz,1H ),6.41(dtd,J=8.4,2.0,1.0Hz,1H),6.11(dt,J=7.5,2.0Hz,1H),5.26(dd,J=4.8,3.2Hz,1H),4.54(s,0H), 4.36-4.23(m,4H),4.05(s,1H),3.72(td,J=4.9,2.8Hz,2H),3.72-3.65(m,1H),3.68-3.60(m,6H),3.63-3. 53(m,4H),3.42(dddd,J=12.8,4.8,3.6,0.8Hz,1H),3.35(tt,J=8.1,1.8Hz,1H),3.24(td,J=5.9,4.8Hz,2H ),3.15(p,J=6.5Hz,1H),2.93-2.78(m,2H),2.73-2.65(m,1H),2.42(d,J=5.3Hz,3H),2.29-2.20(m,1H),2. 20-2.03(m,6H),1.95(ddd,J=12.5,7.8,5.4Hz,1H),1.77(ddd,J=12.5,7.9,5.3Hz,1H),1.59(dddd,J=11.2 ,8.6,5.7,4.0Hz,2H),1.55-1.45(m,3H),1.48-1.23(m,12H),0.89(d,J=1.5Hz,3H),0.84(d,J=1.7Hz,3H). 13C NMR(125MHz,Chloroform-d)δ187.05,172.67,171.75,171.59,170.54,167.73,165.09,164.08,156.92,156.71,151 .55,140.80,135.49,134.65,134.27,133.57,131.03,130.73,129.85,128.42,123.80,122.94,110.16,94.22,84.8 9,70.59,70.30,69.67,69.57,69.54,69.48,67.03,66.20,61.01,59.05,55.83,54.31,48.47,48.16,47.45,40.60,38.58,37.43,36.55,34.10,34.03,33.33,29.89,29.01,26.29,26.10,25.50,24.97,17.21,14.28; This indicates that the synthetic compound MJ-16.

[0150] Embodiment 22:

[0151] In this example, the target compounds MJ-1 to MJ-16 prepared in the above examples were tested for biological activity. Specifically, the in vitro growth inhibition activity of breast cancer regeneration cells (TRCs) was studied by culturing human breast cancer MCF-7 cells for 5 days in 3D fibrin glue (90Pa) and DMEM medium containing 10% fetal bovine serum, and the desired melanoma tumor regeneration cells were cultured and screened (Nat. Mater. 2012, 11, 734). Subsequently, collagenase and neutral protease II were added to the culture medium to digest the fibrin glue, thereby freeing the breast cancer tumor regeneration cells, and the obtained breast cancer tumor regeneration cells were transferred to the newly prepared culture medium and resuspended, and kept in a single cell state. The single-cell breast cancer MCF-7 tumor regeneration cells were inoculated into 3D fibrin gel (90Pa) and incubated for 5 days. 0.1% DMSO was used as a negative control, and all-trans retinoic acid (All-trans RA), tazarotene, and WYC-209 were used as positive controls. Drugs were added at a concentration of 10 μM and incubated for 5 days. The tumor clone size was measured and calculated using a fluorescence microscope, and the inhibition rate of each drug on breast cancer MCF-7 tumor regeneration cells was calculated. The test results are shown in Table 1.

[0152] Table 1: Results of the test on the inhibitory activity of breast cancer MCF-7 cells

[0153]

[0154] As can be seen from Table 1, the target compounds MJ-1 to MJ-16 all have a certain degradation effect on retinoic acid receptors, and show a certain inhibitory effect on breast cancer MCF-7 tumor regeneration cells. In addition, some of the RAR PROTAC compounds of the present invention show better inhibitory effects on breast cancer MCF-7 tumor regeneration cells than the commonly used positive drugs all-trans retinoic acid (All-trans RA), tazarotene and WYC-209 on the market.

[0155] In addition, it should be noted that in addition to the above-mentioned target compounds MJ-1 to MJ-16, which showed a certain inhibitory effect on tumor stem cells, RAR PROTAC compounds synthesized using other linker linkers also showed a certain inhibitory effect on tumor stem cells through experiments; for example, linkers can also use linkers with the following structures:

[0156]

[0157] The above examples are merely illustrative of the present invention and do not constitute a limitation on the protection scope of the present invention. All designs that are the same or similar to the present invention fall within the protection scope of the present invention.

Claims

1. A RAR PROTAC compound, characterized in that It has the following general structure: Among them, E3 ligand is selected from IAP ligand One of them.

2. The RAR PROTAC compound according to claim 1, characterized in that The linker is a rigid linker, a flexible linker, or a flexible and rigid combination linker.

3. The RAR PROTAC compound according to claim 2, characterized in that The rigid linking group is selected from one of the following structural formulas: The flexible linking group is selected from one of the following structural formulas: The flexible and rigid combined linking group is selected from one of the following structural formulas: Among them, n=0-10.

4. The RAR PROTAC compound according to claim 1, characterized in that Any one of the following compounds:

5. A method for preparing a RAR PROTAC compound according to any one of claims 1 to 4, characterized in that: The steps include: S1. Synthesis of ethyl 2-((4,4-dimethyl-1-oxobenzothiopyran-6-)ethynyl)-4-hydroxypyrimidine-5-carboxylate, or synthesis of ethyl 2-((4,4-dimethyl-1-oxobenzothiopyran-6-)ethynyl)-4-bromopyrimidine-5-carboxylate; S2. Using 2-((4,4-dimethyl-1-oxobenzothiopyran-6-)ethynyl)-4-hydroxypyrimidine-5-carboxylic acid ethyl ester or 2-((4,4-dimethyl-1-oxobenzothiopyran-6-)ethynyl)-4-bromopyrimidine-5-carboxylic acid ethyl ester synthesized in step S1 to prepare a retinoic acid compound; S3, reacting the E3 ligand with the linker to obtain an intermediate compound; S4. Prepare RAR PROTAC compounds using retinoic acid compounds and intermediate compounds.

6. The method for preparing a RAR PROTAC compound according to claim 5, wherein In step S4, the process of preparing the RAR PROTAC compound from the retinoic acid compound and the intermediate compound is as follows: The retinoic acid compound and the intermediate compound are dissolved in N,N-dimethylformamide, and then sodium triacetylborohydride or HATU is added and stirred at room temperature for reaction; after the reaction is completed, a saturated ammonium chloride solution is added to quench the reaction, and then ethyl acetate and water are added for extraction, the ethyl acetate phase is washed with saturated brine, and the RAR PROTAC compound is obtained after vacuum distillation and plate separation.

7. Use of the RAR PROTAC compound according to any one of claims 1 to 4 and a pharmaceutically acceptable salt thereof in the preparation of an anti-tumor drug.

8. The use according to claim 7, characterized in that The pharmaceutically acceptable salt of the RAR PROTAC compound is a salt formed by a RAR PROTAC compound and an acid selected from any one of the following: Hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, carbonic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, malic acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, succinic acid, fumaric acid, salicylic acid, phenylacetic acid, mandelic acid or ferulic acid.

9. The use according to claim 7, characterized in that The tumor is melanoma, breast cancer or prostate cancer.

10. A pharmaceutical composition, characterized in that The invention comprises the RAR PROTAC compound according to any one of claims 1 to 4, and one or more combinations of pharmaceutically acceptable carriers, excipients, diluents, adjuvants, and vehicles for the RAR PROTAC compound.

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